784 .
NATRONIELLA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
9.5 - 10.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.2 | g | ||
|
|
MgCl2 x 6 H2O | 0.1 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
KCl | 0.2 | g | ||
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NaCl | 16.0 | g | ||
|
|
Trace element solution SL-11 | 1.0 | ml | ||
|
|
Yeast extract | 0.2 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
|
Na2CO3 | 68.0 | g | ||
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NaHCO3 | 38.0 | g | ||
|
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Ethanol absolute | 5.0 | ml | ||
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Wolin's vitamin solution (10x) | 1.0 | ml | ||
|
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Na2S x 9 H2O | 1.0 | g | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients except carbonate, bicarbonate, ethanol, vitamins and sulfide. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve carbonates and sulfide while gassing the head space only. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add ethanol and vitamins from sterile anoxic stock solution prepared under 100% N2 gas. Vitamins are sterilized by filtration. Adjust pH of the complete medium to 9.5 - 10.0, if necessary. | |||||
Trace element solution SL-11 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 5.2 | g | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
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ZnCl2 | 70.0 | mg | ||
|
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MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
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CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
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NiCl2 x 6 H2O | 24.0 | mg | ||
|
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
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Distilled water | 1000.0 | ml | ||
| 1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
Riboflavin | 50 | mg |
|
|
Nicotinic acid | 50 | mg |
|
|
Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |